Disruption Of The Nuclear Matrix Attachment Domain Of The Dna Replication Factor Ciz1 In Common Solid Tumors And Its Potential As A Biomarker

The DNA replication protein Ciz1 promotes initiation of mammalian DNA replication in cooperation with cyclin A-dependent kinase, most likely by delivery cyclin A to sites where cyclin E-dependent pre-replication complex assembly has taken place. Normally, Ciz1 is anchored within nuclear matrix-associated foci that co-localize with sites of DNA replication, but in the absence of anchor domain Ciz1 retains the ability to promote initiation of DNA replication in isolated nuclei so attachment to the nuclear matrix is not essential for function. Here, we show that expression of DNA replication and nuclear matrix anchor domains of Ciz1 are uncoupled and uneven at the transcript level in a panel of common cancer cell lines, giving rise to excess DNA replication domain protein that is not attached to the nuclear matrix. Moreover Ciz1 domain expression is also uncoupled in a wide range of common solid tumours, including breast and lung, so that anchor domain transcript is elevated over control tissues, and exceeds replication domain in all primary stage I, II and III tumours tested. Notably, more than half of all stage IV tumours tested resemble established cell lines, having more replication domain than anchor domain. In cell based assays, recombinant anchor domain protein interferes with attachment of endogenous Ciz1 to the nuclear matrix, revealing a dominant negative effect that also impacts on nuclear matrix-recruitment of a key component of the pre-replication complex, Cdc6. This shows that Ciz1 normally plays a role in localizing Cdc6 to the nuclear matrix and suggests that cancer-associated uncoupled expression influences recruitment of mammalian pre-replication complexes to the nuclear matrix. These findings implicate spatially unconstrained DNA replication as a source of nuclear disorder in cancer cells. We identified a variant Ciz1 isoform with alterations in nuclear matrix attachment domain and tumour-restricted expression. Variant Ciz1 is elevated at the transcript and protein level with high frequency in small cell lung tumours, and with lower frequency in a range of other tumour types. Selective suppression of variant Ciz1 expression using inducible shRNA restricts proliferation of lung cancer cells that express it in vitro and in vivo, identifying a novel, exploitable therapeutic target with potential application in the treatment of lung cancer. Recent evidence will also be presented on the application of variant Ciz1 as a serum biomarker. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3022. doi:10.1158/1538-7445.AM2011-3022